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In order to study the impact of biomass from different filter depths on biotreatment performance, a bench-scale study was undertaken in which the biologically active sand media from the pilot plant filter was broken down into four segments and loaded with pre-filtered, ozonated water. The objectives of this study were to investigate the impact of contact time on biofiltration performance and to determine the removal capacity of biomass at different filter depths. The organic matter characterized included dissolved organic carbon (DOC); biodegradable components AOC and BDOC; specific ozone DBP aldehydes and ketoacids; and chlorination DBP precursors.